Patent AU-698442-B2 (AU)
Title: Modified oligonucleotides, their preparation and their use
Inventor: SEELA FRANK
IPC: C12Q1/68
Filed: 1995-05-01
Granted: 1998-10-29
Abstract: Oligonucleotides (I) containing a triazolopyrimidine (8-azaguanine) residue, and their salts, are new. Oligonucleotides of formula (I) containing a triazolopyrimidine (8-azaguanine) residue, and their salts, are new. R1>hydrogen, 1-18C alkyl, 2-18C alkenyl or alkynyl, 2-18C alkylcarbonyl, 3-19C alkenyl- or alkynyl-carbonyl, (6-14C aryl)(1-8C)alkyl, standard protecting groups or -P(=W)(Z)(Z'); R1a>hydrogen, 1-18C alkyl, 2-18C alkenyl or alkynyl, 2-18C alkylcarbonyl, 3-19C alkenyl- or alkynyl-carbonyl, (6-14C aryl)(1-8C)alkyl or -P(=W)(Z)(X); R2>hydrogen, hydroxy, 1-18C alkoxy, 1-6C alkenyloxy (particularly allyloxy), halo, azido or amino; X : hydroxy or mercapto; U : hydroxy, mercapto, SeH, 1-18C alkyl or alkoxy, 6-20C aryl, (6-14C)aryl(1-8C)alkyl, NHR3>, NR3>R4> or -(OCH2CH2)pO(CH2)qCH2R5> (III); R3>1-18C alkyl, 6-20C aryl, (6-14C)aryl(1-8C)alkyl, 2-(CH2)c-[NH(CH2)c]d-NR6>R6>; c : 2-6; d : 0-6; R6>hydrogen, 1-6C alkyl or 1-4C alkoxy(1-6C)alkyl; R4>1-18C alkyl, 6-20C aryl or 6-10C aryl(1-8C)alkyl, or NR3>R4> complete a 5- or 6-membered heterocycle, optionally containing another O, S or N; p : 1-100; q : 0-22; R5>hydrogen or functional group, e.g. hydroxy, amino, 1-18C alkylamino, carboxy, aminocarbonyl, (1-4C)alkoxycarbonyl or halo; Z, Z' : hydroxy, mercapto, SeH, 1-22C alkoxy, -O(CH2)b-NR6>R7>, 1-18C alkyl, 6-20C aryl, or 6-14C aryl-(1-8C)alkyl or alkoxy (where aryl includes heteroaryl and aryl and which may be substituted by 1-3 carboxy, amino, nitro, 1-4C alkylamino, 1-6C alkoxy, OH, halo or cyano), 1-18C alkylthio, NHR3>, NR3>R4>, group (III), or a group that promotes intracellular uptake; is used to label a DNA probe or, when (I) is hybridized to target nucleic acid causes bonding, crosslinking or cleavage, and the curly brackets indicate that R2> and the adjacent phosphoryl group have 2'.3' or 3',2' orientation, and where each nucleotide has D or L configuration, with base B in alpha or beta position, and each a standard base, e.g. adenine, cytosine, thymine, guanine, uracil, hypoxanthine, or an unnatural base such as (8-aza)purine, 2,6-diaminopurine, 7-deaza-adenine, 7-deazaguanine, N4>N4>-ethanocytosine, N6>N6>-ethano-2,6-diaminopurine, pseudoisocytosine, 5-methylcytosine, 5-fluorouracil, 5-(3-6C alkynyl)- uracil or cytosine or their prodrug forms; b : 1-6; R7>1-6C alkyl or R6> and R7> together complete a 3-6 membered ring; a : O or CH2; n : 3-99; W : O, S, Se; V : O, S or NH; Y : O, S, NH or CH2; Y' : O, S, N (CH2)m or V(CH2)m; m : 1-18; at least one B' : a group of formula (a); and E, F'' : H, OH or NH2. Independent claims are also included for the following: (1) method for preparing (I) by successive coupling of nucleotides to an appropriately derivatized carrier or to a growing oligomer chain; (2) nucleotide monomers of formula (V); (3) method for preparing (V) by converting a nucleoside (protected on hydroxy and amino) to its phosphonate or phosphoramidate; and (4) compounds (VI). In (V): Yb>O, S or CH2; R1'>standard protecting group; R1b>-P(U)(Q) or -P(O)(OH)(O->); R7>2-cyanoethyl; R2b>hydrogen, OR12>, 1-18C alkoxy, 1-6C alkenyloxy (particularly allyloxy), halo, azido or NR10>R11>; U : OR7> or SR7>; Q : NR8>R9>; R8>, R9>1-6C alkyl (particularly Et or isopropyl) or together complete a 5-9 membered heterocycle that may include another O, S or N, especially N-azaundecanyl, pyrrolidino or morpholino; E', F' : hydrogen, hydroxy or NR10>R11>; R10>, R11>hydrogen or standard protecting groups, or together complete a protecting group; R12>standard hydroxy protecting group, e.g. tert-butyl-dimethylsilyl, triisopropylsilyl, o- or p-nitrobenzyl, tris(1-methylethyl)silyl or 2-fluorophenyl-4-methoxypiperidin-4-yl; and R8>, R9>1-6C alkyl or together complete a heterocycle. In (VI): U' : U'' = U''' = hydroxy or mercapto; e, f : 0 or 1; R13>hydrogen or hydroxy; and provided that the compound with U', U'', U''', R13> and F : hydroxy, E = amino and e = f= 1 is excluded. [Image] [Image] [Image] ACTIVITY : Anti-HIV; Virucide; Cytostatic; Vasotropic; Antiinflammatory; Hepatotropic. Test methods are described but no results are given. MECHANISM OF ACTION : Inhibition of gene expression.